茶树miR164a及其靶基因的鉴定与表达分析

孔雷; 朱向向; 王屹玮; 谢小芳; 江昌俊; 李叶云

doi:10.13305/j.cnki.jts.2018.06.001

茶叶科学 >

2018 , Vol. 38 >Issue 6: 547 - 558

DOI: https://doi.org/10.13305/j.cnki.jts.2018.06.001

茶树miR164a及其靶基因的鉴定与表达分析

孔雷 ,
朱向向 ,
王屹玮 ,
谢小芳 ,
江昌俊 ,
李叶云

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安徽农业大学茶树生物学与资源利用国家重点实验室,安徽合肥,230036

孔雷,男,硕士研究生,主要从事茶树栽培与育种,E-mail：13655518357@163.com。

收稿日期: 2018-03-07

修回日期: 2018-03-29

网络出版日期: 2019-12-15

基金资助

国家自然科学基金（31270729）

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Identification and Expression Analysis of Tea Plant (Camellia sinesis) miR164a and Its Target Gene

KONG Lei ,
ZHU Xiangxiang ,
WANG Yiwei ,
XIE Xiaofang ,
JIANG Changjun ,
LI Yeyun

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State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei 230036, China

Received date: 2018-03-07

Revised date: 2018-03-29

Online published: 2019-12-15

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摘要

植物miR164通过负调控靶基因NAC转录因子（NAC transcription factors）参与植物生长发育调控和逆境胁迫等复杂的生理过程。利用茶树microRNA高通量测序和降解组测序,对茶树miR164a及其靶基因NAC1进行了克隆、序列分析与鉴定,并对其在叶片发育、高温和低温胁迫中表达模式进行了研究。茶树miR164a前体序列长度为126βbp,成熟序列21βbp。CsNAC1 ORF框长度912βbp,编码303个氨基酸,在CsNAC1蛋白的N端有典型NAM结构域,C端有Csn-miR164a识别位点。RLM-5´RACE验证CsNAC1为茶树miR164a的靶基因。荧光定量表达分析表明,Csn-miR164a在芽中表达量最高,第7叶中最低。在高温和低温胁迫下,Csn-miR164a都下调表达,而CsNAC1表达水平上升,CsNAC1与Csn-miR164a呈现负相关。研究表明茶树miR164a及其靶基因CsNAC1可能与茶树生长及抗逆相关。

关键词： 茶树; 靶基因; 温度胁迫; miR164a; NAC

本文引用格式

孔雷 , 朱向向 , 王屹玮 , 谢小芳 , 江昌俊 , 李叶云 . 茶树miR164a及其靶基因的鉴定与表达分析[J]. 茶叶科学, 2018 , 38(6) : 547 -558 . DOI: 10.13305/j.cnki.jts.2018.06.001

Abstract

Plant miR164 is involved in complex physiological processes such as plant growth and development regulation and stress responses by negatively regulating the target gene NAC transcription factors. The identification, cloning and sequence analysis of miR164a and its target gene NAC1 in tea plant were carried out by high-throughput sequencing of microRNA and degradome analysis. The expression patterns of Csn-miR164a and its target gene CsNAC1 during leaf development and under high or low temperature stresses were also studied. The length of Csn-miR164a precursor is 126βbp, and the mature sequence is 21βbp. The CsNAC1 ORF is 912βbp, encoding 303 amino acids. There are a typical NAM domain at the N terminus of CsNAC1 protein and a Csn-miR164a recognition site at the C-terminus. Quantitative real-time PCR analysis showed that the expression of Csn-miR164a was the highest in the bud and the lowest in the seventh leaves. Under high and low temperature treatments, the expression of Csn-miR164a was down regulated, while the expression level of CsNAC1 increased. The expression of Csn-miR164a was negatively correlatied with the expression of the target gene CsNAC1. The results revealed that Csn-miR164a and its target gene CsNAC1 might be closely related to the growth and stress responses in tea plant.

Key words： Camellia sinensis; target gene; temperature stress; miR164a; NAC

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